Plasmid-host systems

Abstract

Plasmid-host systems are provided that allow for maintenance of the plasmid independent of antibiotic selection.

Description

[0001] nvoernnoer:oecsse e ao..: - - - - ,,se a oncuaneon, cam,s.r.orrorom..rovsonaaen caono. eruar, vsonaaen caono. ,,e on an..rovso,na..aeronvsonacaaoenno.caono. ,,e oncoer,,sncororae reerenceerenns enre.,,e onuus, , eac o wc ncororaee reueenrceencesnns eansreen sume eecroncan orma ansere _.xm ans,e.eesn sze c.o, creae onuus, ,s name correceneacsmee-cass aenneneeerevveerasrovenromsnneneera acaonso raonaeraeucansmon anreamenca on ceocousseases an cancer. roaaonn aaceraosusuascerca co an a euamrrses a un essenaor secca,raonaeraeucasm comrses: an oronc oranscron un.ore or ressanceo an anoc an aransene w seuences nece recaon aene e.., enancers,romoers,oaenaon seuences ssarors exresson srea oeu acnraoco rneassaanscmesenerseseon o seevreraac ceo, rn a acne orrns.e erose ss,. concer ons enxesra reoanrne os’saceunromne.rocuecss, s ereear corouu o wcc noe manu ausre w sreanms oer aucmcanenao re aeanse,orouernnaeeaansmo nvoerneoceo.: - - oee sno nraera:e oss m r e aecs as. nancuemaenne asnn anom naauraaceraouaons,osn ecooca rss anare, ea.econ, sucraonaasms are reave euar aoncraasnmsec sozne eercmeena aecs nucec aceveraceraransormaon an reance oneseenes maenac.ossrue,sr waonaasmasmosresenroucon caenes as anoc uses no aeuae manane, comcasns orens maanuacu srenecvroecersessure reucnes s an asms we.rn cave anoc ressance-ree seecon ssems exs.or exame,e ressance seecoonn ssoneams o rren on o ree sucareosnson an oe asmsree o anocno an essena cromosomaaceraene.easm an asm conera nnonsense muaonnrouce ansers a ceran amno acnsea oermnanransaon.e anos am srearn ssuorresrosor o s an saveeen moeo carr an amer so aaon can one comemene a coonn an essenaenea a s wereesne corore osveorncomne s aun arresnsonre auxo.ro, cause nroucon o rens asmanecre meunaeoonr an sonee acceornneeaceaseenear, reacne anoc orn o recaona reu arems aenr su. coressoors sra.na exs ursee an-erve conona roen encoe aene sses arans-acn πnaor asms user a sa cone nraenoe cromosome.snroucenoemaeeneum, ceornseu oenrn oea rencoa aon.me anme aeurx mouraon sran.an,e comemenaon suressons ac oc oneasmacone. evenroucon o a suressor nvoernnoer:oecsseo.: - -neenen oa e a ans. neoece sesec aona,sm an- wosere snsemeaasm aowsor manenance oeasm acone. as a comarave smaasm recomnanno osne ce as execr,es asnro aaereoroco reocuosmonnanuos cesrove,eroaroc comrsn areneaas unerone arames mcueoaoen, comerrasmnes an o men reanramen aremaure so coonownsrea uaon resun neo a mne reanrame,erameserene oera rameseromroeenre anur ceor cnoesor aruncae recaonnaorroen.n one asec,enaorroen.n one asec,ore a rreoacraooncn recoamornroen, e.., auen recaon eerooousonuceoe comrsn an oen reanramea cnomorssn ce aurreenr exresses a oera neo aromoer an conor a re e,erene recaonnaorr caonnaorroen e.., auen renes annucoeerno.mno oenr.e ans oence, aseecro,moeras oeraneoeramese aeerooousonuceoe comrsn a conoenaro easrseonca receomnanos ce exresses mue aamone.r mnu oaneon as coemc,rsees aea sno m cuoaononn comacer oses a an amneer m coumarosnn.n a oene asaenc, recaonna anso caera ase oec a,c avaaosmnserrouvne,easme. comrsn: an orn onaorroe cae recaonnaorroen,e recaon erenne c,o ore ao cromneaoranmesereoer aen see,ece recaonnaorroen coeor onuceoe oera neo reuaor seuencesa ane m eaxrreerss anen aeereoroaorooucs nvorneoceo.: - - reecoe ermnor oon:aoness usoo es a. nu cece.one c oonmer assnec, a coenroonaaro ecsse rnecoamennaen comos ce exresses a muaon, ane seecae marer comrses a srucuraene seuence o ars sun areean e suressor comrsn an ancoon c ssor , essenaene. aae o comemenne conona onuceoe conm ornes asec,eroaroc recomnanos ce exresses aeerooous e seecae marer conm ar csoesn auona se essenaene comrsn aean muaon, an ene. uenceeneo comemene conona essena comrsn:n an couevra asnec,e a mroeaosroveor mucaon oeasm,e meo er roc recomnanos ce nroucneasmno eno caarrreo ocu recnom a mnanos ce seecneasmrome ce cuure,e seecon row an cuvaenume sereeec oe a c neusr.en vaoreroaroc recomnanos ce’s e meo cno amnorsern as:ec, a meosroveorroucn a recomnanroen oneres, eos ce comrsn: m aueeroonouseoasnmuceoera cnosmorrmsn aos ce easm,o aromoer, an aeerooousonuceoe comrsn an con aroneane o esesreanane cuvaneos cen a cuure meum aown e eneneres an xresson oe recomnanroen oransorme urne recomnanroen oneresrome cuure meum orrome no asno ce. comrsnransecern aseec,- ace me woseroavsme.orrearn a moe-ce,e meo nvoernnoer:oecsse e ao..: - - comrsnnra annsoecern as aec eu, aar moecooss cerov weorereasamrn, v wraere vnecores,ee meo onuceoe oeasm comrsesenes encon eemens reureorrouerooous vecorsne cn vraranscronn. a enuoarero ascecos, a c mee.osroveor use oeasm as aemaeorn vro seccaon,e are ac ucsoem maenrenouursesr,a we vacrou asre exnacmoreor aaseen an consue aar oe onuceoeu,reeo snuocweso ane exameasma “cms.a”asm comrsn a seuence conor a recomnanr coomenr osn: an orn o recaon a nuceoe e recomnanroen oneres neres an exresson casseeor exresson o ,e suressor comnr asnos ce an a srucuraene seuence o a suressor essenaene comrsn aeson comrs ann a ancoon caae oarn w a conona acone conro conanne orn o reecaaonn a mnua aon.ure sows a sanar cassee.ure sows a secon sanaracone n anoc ressance exresson smaroe s asm conro w a smaer sze, more uzre oema exame sownnure. uorescenve ceese-, u soonwrane oave cee, ercenuorescen, an ure reaveoe conros ssoewcnonn our ceess w anm.aasms oee sownn nvoernnoer:oecsse eo.: - - oee souwrne as. n aunre s reoawveoea caon ororsan ssoewcnonn our ceess w anomaasms os-eecroor r,ours sownnurua ereon. s reoawsveoe caoan oror sanoswencnon oure ces, wourmsaos-easemcrs oee ures an sow exameromoer arcecures o annuceoorraenon. ure sowse ceens aane cuur e.ransorm es oree coonesorme sranearnn a,nemra aa,asm oee sownnureno “ ,” wcs an. co escre wu rreesec soow as mueaea asmsenee ane ar m mua aeerrouresne o.rowor eac cuure escre wure sowsu arearo.sees o sames oasms exracerome cuures u rerseec soowsure. cuuresorme ransorme ceens aerours orown mnma mea aane erenos srans, eac o wnc asm ana. caosm oee sownnure noour anree o wc“ ,” “ ,” an “ sran”earena,ner aa, a muae arene, ure sowse ceens aerours orro aw munaeurar-eraenne. or suosw annnnucunr aen aane cuuresorme ransormn amaasm oee . ueren soowse oasrme.ser m o eac cuureescrenures an nvoernnoer:oecsseo.: - - escre wu e rree a. sec soowsu arearosees o sames oasms exracerome cuures escre wu rreesec soows aar.osees o sames oasms exracerome cuures resunromu .rraensorm sowsur aee.a sows uncu anac-eseasmrearaons wcs an n amaasm oee sownnureno “ ,” u core sra snowesarne,mnear aesa o, a meuaaesm arrene an ar- ene. ure sows aea sows uncu an uneeasraeon raesmerrereoanraonusr ree. romransormn one oeemaes ree sun co sran sma rreonureno “ ,” wcs an. urreso an, exce sowea srenec aen resunsesr oon excnsne ar m-onene. asmrearaons reerreonure an usne monomeromaneroanra ons oone oe an aa rmonuresur aen so wwn.se r eexsuonusc oeasreea orn res servceraon o enzemeascmo.rearaonseneraes exresseroumrere seowserean.ro cmooerarsn,sourm naense arre o oseer mveua wnenr ar muanene ene we,uenruncae, are aeoacaeasm recaon oeene anmaeasrm. sranransormuere a sowassm exa cmomersn-aeee resus oroen exressonrome romoer, a orn o recaon, an a srucunrae enecnoe senueeroen, aa nuce eassor seeconse ar* amer so muannncee o a sureessnoorme, w, wceres nvorne oce o.: - - comeneomr:eneess e a. sranurraense su sorm aenressor a saosw. m exa cmomersn-aee reensue esn ocornoene exrroesesnonromenuceromoer, eer a or orn o reca , an - a sures on, an a srucuraene seuence o enome, wsorcs c,om weermeeneeassoer s sueecreosnse ar* amer so muanne ure s ssowemss exaemscreeunoresces uno rcr ees res. romeos-asm ,us o, anroen. aser- exresson conex cearse ocaees s oneurwarseo.rmsor “a e,x”am “ean,n e”, r ao enns er “encee”on “caueeneu oran reeerences unesse ura o res” mancue anercaneaeernmess o er seuncere n an ass. en “ccoomnneacsse,”rec “a asa wcee, a”s “nnreec, c”on annec “ocno,n auaae” are use or conuaon unesse conex cear caes oerwse. ere a rane o vcamueesn,s rnae, eacnervennneer vaue, an eacr ece, ms aconereo,eweene rece uer anower e u oer aan ranoweers amsos secca scose, aon w eac suraneeween suc vaues. rane, an eac rane wer oe e ane rra,n nee caenrneenen encuen or excuerome ere a vaueen scusse asnerenm, ors,oor exammse arencues aso encomasse. a a concenraon orom o , or weree o an au weoeurse s ao cuomoonen caneresen ,osenerenms are secca scose. ere a vaues excn c raenc raen,e varuoemsao nvoerneoceo.: - - aree “n sao cooru: e.”essa e a erse,. w a comnnaones samsceouan or amoun ase rece vaue are aso wn comnao se, eac su-comnaon oe eemens oa eemens arnens avsou saeccsacosescose.onverse, wereeren eemens orrous osscose asavn aura o, c aomernanaveosn,s exaereo are asoscose.ere an eemen sn orn an comnaon we oer aernavmes aerse ans woecre eacs acoersnaves excue eemen canave suc excusons, an a comnaons o eemensavn suec m exocreusoan one eresco ns are meann ansees.sene oerwseeren, aecnca an scencermsavee same conars o commcorono uoner asnoooe ocune o ornar sndne reevan ar.neon, e a., , anae aram,e aroo, .,one anons,ewor ,ro areronsconar ooo,arererenna,, oerwsevnec oanee o, nu scec w ac as areeneraconar o man oeerms useeren.ness seuences are wreneo rn am wnroeon caeroxo r orennao’n.o’ orenaon an amno ac eow are moreuene reer eermsenemmeae n “amer nonsen enceoe seccaon as a woe. n wc aoee canssee mrmuanaoen” orrem “aamuer muaon” or “amer coon”s a muaon susuona convers a coon secn anr aem.nom acer mnuoaons, ar wece re ssun o aaseermnaon. as cano reconzene “amer suressor ”s a wose ancoonaseen canen orer amno ac aa unouse noonnsnensee cooone’-e ca-n’.amerne m annser an aernave nvoernnoer:oecseo.: - - a cannorns o “ eauu a cx. eo ar nource”n s vraanosr aroacwera,ueas,rooas, or mammaanos ce sran unaeo sur eoenec muaons.s a resu,e srans conras,roovrvoenc m mecroaoracannsms caan s secc nuren unesssrove exerna.n rowrom m nesze a oe nurensa are reureorer “cnonmao mneaum ess woue aon o suemens. oerovso enaene”s aosneceneas essenane asence en “ oco an nuurreano exoenous. wneonuceoe ann”e o or aer aonnucreecoone o meaonsse eeemuncona seuence eemens “conservave muaon”se resu o a msseens. suson o a smaru noenca am nse muaonaeasoe consss o a muaona susana reannso aecu.ncosn u oseee orrenn,a a conservave muaon eerms “corresonnransos , unmuaeroen.n on” an “corresonnransosase” are useo s ccoarrees ano ancv reaonseween aransosase an aransoson.ransosaseransoses mssense musa unsoeranesreons,o an. “ean muaon” reerso a nonsense muaon or aean a co eenrm, sse “uence seeeuenocne,, o”r “ seuen sceeunesnecre,o”n o.r “onuceoe se ennuoous a nucec ac seuence uence” reeron .e seuence cane an oonuceoe oo nuceoes eue-rmen “exreensosmonc c soenusernce oousans orunres oousans oasears. , suc as,or exame, a consrucuc”a me caonnsa anns aoenasuc oenoeesneoranscre an oeraneo aownsreamene, con reon, oronuceoereo smeoueernceaosr e oxra mmae,e a nvoernnoeoceo.: - - c moecu or: ereesnso em ac. ramena encoes aoee orroen, or an eecor an,or exame,a ann anaseanmseer, a,oruee-xs-roarnmen ,, roro ezxame, an arca seece comrsn a sem-oo orarns, or a- me, an moecue or an .n “exresson v ner or mu-ners or a mcro, romoera cane oeraneceor”o o ar s “eecxonressoonnucasemo”e.s araonnuceoe comrsn a oe exresson consrucn secon orransormaon moecue o a recen ce aowse ceo exress an eecor mae a,eoneecae e,n orneroereen enacsome, vre exresson consruc.n exresson consruc erverom,or exame, aaceroaeu,s, a reecnoomvrunsa,n a vernuos, or an arca cromosomeenvrus,oxvrus, oreresvrus.uc exresson vecors cannc-ausseoc sae vrus, rerovrus, vruses, oraes.uc vecorsncue cromosoma, esoma, an verusu-enecrevsero vmecacera, exame, vecorserveromaceraasms, ors,or cromosoma eemen aceroaes,eas esomes,eas erveromasms, a ann varcuseerso, veacoersenerevce ereomm comnaonsereo, suc asose exresson consruc ca ens, cosms, anaems.n “e ne recaen avn ce, or cane mae sneca.eerms exrernessoonem coonnssruc,” “exresson vecor,” “vecor,” an “asm” are usenercaneaneneomraeenveen aoncoa aonar ocuaernveen oon exnre aenera,usrave sense, an are no eerm “exressonoee” means asson consruc. exresson con oee encoe aene on an esreurmc. useorouce aen “eexroresuscon en scoseem” m aeanos annuceno vveo. orn vroooca ssemas nvoernnoer:oecsseo.: - - ownsream re “ e aram a. nesr muaon” occurs wen aasenseron oreeon causes a sne comrsn “eeneenreanamsoeer. seseramn”s reerreerso w a vcecosr o croeneranser vecor,.e., aonuceoe onuceoe” or “en neno a vecora “eneranser eauresoacaeeerraoncsesesr o consenruec”ra.neneranser ssem ma aso comrse oer comrse a vecor an a or vraacans mer.xoorr en eaxamne a, arsenoenrauncseer ssem ma ce, or ma comrse aonuceoeancue oeo ener a seuence en s aransoson an a secononuceoeransoson.coenr aan csororessaseosnnransosaseo enanceroucveenomcneraon oe nucec ac moecue or on aenrenra nnusceoscon acs o m aoeecneranser ssem mae one same ssem maero ues.eransosase o aeneranser wo eevmeens as a areo “enuceoe or as aoee. exame, a nucec ac seuence eenrcooonous a”rooe onne annoeeor no naura assocae.or romoer oerana wc naurarves exresson oe areoeeron.oouseerroomoooeurs m neans a acane ransoson ens ornvereermna re ucec nucec ac no naura eas“”s means aeerooous encon aoee oeranaen aransosoesasrea,nsncosuonn en ans a onros, suc as a nucec ac nucec acseerooouso a ce no nau eav or can. ceun aerenocao raounne ce or nauraounne escre. ne.., esoma orerenenomcocaonaneocaon o a nucec acee.rm “ “ooss”” mnecaunse asnroroararooc ocr o eru euararoocc or oranansmsmsaa c caannee ae rneeccaen nvorneoceo.: - - ene annn ueo saer “,r:e oso.esso er ,”r e ax. namareso or saucoss, seeanas e a.,oecuaronn.aoraorerm oraor,ornaror,...s useeren,e n “ “onsro cne”,” “os ssem,” an “exressonos” cane usenercanea. ena r s a nuceoe seuence wn aeneas no exresse or oeraven aaseenn “o esouac ere. r”emooveerome orso nanuurcaeo enver monemanesn aoee oronuceoeecnues, or cemca or enzmaca snesze.oe,eros ourceo unsucneo receomnan ure,as, essenareerom an oeroe s mae ceuarroucs o e oronuceoe an assocae eren amn “mssre onseer mmuauron”es o.ccurs wen aon muaonrouces a coonaes a susuon o “on aocn-c.onservave muaon”se resu o a mssense muaonaeasoeeano ar “un a nocnass eesnms aen maru am oassonno” a oc ncoc. nurusn wcen aon muaonrouces aremaure so coon eerms “nuceose” an “nucoenoaroen. enownurne anrmneases,u aso oe”ernceueroeccosce maoseesa conan no on uc mocaonsncue meaeurnes orrmnes, acaeesaurnaevse oreenr mmonees. or oereerocces.oe nuceoses or nuc , suar mo eoes can asoncue mocaons one orsunceon,aozre exa ams ane e, weere aroxrous reace w aaoen, an aacrou, encomass nuceoses an nurc, aeno amesn.e, oree.eerm “nuceoc un”sneneo nvoernnoer:oecseo.: - - e m ons “ eoec a.reerm mnuaaoron c”oson a nonse,nseea msuoaoneoarm,aueono o aerruenmcaauere, aossearanc neonn uncona,r - m ono e “eon. eram mnauoaro cno”osn a nonse,nseea msuaooneaor,muaeooneremaure aearancene uncona,r o aruncae,oss non- noen. ransaeno a “moenno r aecasn, crame” or “” means aoron o aonuceoea, wen rous oreeasears, wonca mnesa nnos so coons.eenec coe reas seuencesn sxosse reanrames-reeneaor awaoruer-escraonne moecue can rean an o ca asoncues ann anreene reverse.n eerm “o aon coon a wcransaon ma sar. a one seuence moeraeseneea”v reersounconanaeeweenwo seuences suc comrsn a nucec ac exresson coornr oo seeu oener.or exame, arsonuceoe enancer, or arra oranscronacornn sesc aens auc as aromoer, seuence, ne ersonuceoe aecsranscron a snecoonrroansnaucoeno oe aree o seera onuceoe.mar, ars amno ac seuen con suce ce comrsn a secreon sna,.e., a amnou acaro secaueznaon sna, an a secon amno ac seuence are oerane ers suceuaro ce causese secon amno ac seuenceoe secree orocazeo a ncaon. cromosome, “oasrmn, o or r verusc.aon”s a seuence o a wc recaonsnae on a nvoernnoer:oecsseo.: - - unconore e anoe arm. er “aosmnuc”e roeeres.o aoro exnaumceoe,easmass useoerorm a “carrn” onuceoeoeroaae wn avn are oen useo aow a everno a ce, oro aow aonuceoe ceo,eon aeowra aeononuceeoenoemoceac oa ae ceor asm maurer comrse aonauncona eemens, .ransoson. suc as,or exame, a comrsnee orermn “ oa resmcaoacneo.n.e,” or “ ora ocrone” r aenerseo seeecaoenuceoe semen eerms “onuceoe,” “oonuceoe,” “ marer. moecue,” an “ene” a nucec ac,” “nucec acen, an ma comrrsee us reonnuecrecoanees,aeoxo rreero aomercorm o nuceoes o an ereo.eseerms reer onoermar srucuroenu oceoe meos,ec aunae.osereo, or mxuresre-,oue-, an sne-srane , us,eermsncue eerms a as we asre-,oue-, an sne-srane . orms oesoo encnoumceaoss me.ooere,oarr excaumare,e aeaon anor can, an unmoe “nucec ac” an “nucec ac moecue”ncuermos “eooxnruconeouceeo,” “eoonuceoe,” eox--rose,oronuceoesconan sconann- s , an m, weer sce or unscn-rose,ncun , r, , or-cose o aurne orrmneasee, a,n an o oeerroem oerso connuacneno neonnaucs an- acones,or exame,oameor exame,ee n eoc omoronocommerca avaaer ucec acs“s” an omers, an oer snec seuence-seocmc neucnec- acraso,nmc.,orvas,re., aseuene conan nuceoasesn a conuraona aowsorasearner asnrovasen sacan,e suocm aserss nvoernnoeoceo.: - - “ouonr nun:ess e ceoe a an..eres nonenesnconnen eweeneerms are usenerc,”an “oeaonuceeroen.e,” “nucec ac,” an “nucec ac moecue,” aneseerms ooeoxronuceoe’’oseseorearmmsanecs,u’e-,-oar ex-asmuse,ue’-eox-,’,ou’-, sne-srane , as we asoue- an sne-sran e- an or examerseween an orewee , an rsereoncun wnownes o mocaons,or exame,aes, aenaons, “ acnas,” su orsuo,n an o a a nsuocnecouee or e axnam anaeo, m,enernucoesooen maoescaons suc as,or exame,ose w uncarenaes w neave carenaes,oro esxamoree,serso,soosroooraames,aeso,s caroamaes, oree e, an w osve carenaesor exame rooaes, ore amnoa osoresers,ose c , amnoa osoramaes, ncun enzmesor exame, nucoenaasensn,oxenns,a ann moees, suc as,or exame,roens ee,ose wnercaaorsor exame, acrne,osoersa,e sn,n oareeees,,oose- c-onsane, or ceaeso,or exame, meas, raoacve meas, nn conann aaors, oron, oxave meas, oree,ose ee, as we as unmoose weo mrmos oeenoaensucoero exaem or oe, aa anomerc nucec acs, or “on muaon” occursn a nucec ac se onuceoe. eee, or ca uence wen a sneases ae, oerane “nre noum. coeecr” ac me manose acu nuec.ec arcomo seeruence sucenorecranscron o an conro eemensor exame, enancersa are s cuanceen useo reoneererro wmo oerranscron exresson conroaen a cee-secc,ssue-secc, oremora-secerc- meaennneer,n orenae nvoernnoeoceo.: - - a wrennur ac:e nness e ron. e ax. ner onnae a ssneacs, ore areonmso seurc ma eemeens mae wne’ reon o aene or or exame, a c, a oeraneo a nucec ac seuence, enae exresson oe nuencee sce aucenc see,u oern acne eecor con seuence,n suc a wa asono wc a seece nucec ac seuenceo, oer aranrsocmroeer csanroeven an exresson cassee “recaonnaorroen”s aro convenennsere. wn an orn o reca ena reconzes secc seuences eerm “seecoan aen maraceraes recaon oe . or o ” means aonuceoe semena aows oneo seec can ern acoaens an a a mcovecu,e s or a cea conans, oen unerarcuar conons.ese marers ese marers canrovuce a as,nun no smeeoro,ro,uecones o,roe,n as,ene, or aroen, or comouns, or comosons.xam oranc an oranc semen es o seecae marersncue,u are nomeo: e.., anoscsa encoe sreomucenssaaro evncoee resrsoanuce aans oerwseoxc comouns recen cee.., enes, a csa are oerwseacnne a sures uxoroc marers semensa encoeroucs rea ensee ace.v., oen aoenec mroarucers suc as seeam-eanascoas encoeroucsa cane roens semensanroucsa ase, , an ce surace anoruncon se are oerwseermenao ce survva escrenos.- aovemeen.s., aan oseenrswes oenonuceoe acesv o an o seemens seamenns ursoeucossoaae m aoesr ae su msoraeceuee.e.. re.srcon enonuceases semensa cane encoe a secc nuceoe seuence seac ccanroee onernwsne n sones-unconae s.e.,meonrsa nvorneoceo.: - - ameno reccr oa: roe nnss o e rec s au. coonuear soennss ov moecues anor semens, wc wen asen, euenceen caneeoerarcuar comouns. , , an nemnescons annenne secsuenocewsa ursena acaoremes,ea susec as enecsomuerrou, cencer.,ason,s .nsecon, ane ., usneauaarameers, or over a comarson wesn aown.mener.e., resunneesercenae o seuence smar oma ane seuences over ace wnnaoew o o se coumencaerens cacuae comarnwo a wceenca resues occurno seuencessoon,eeerme nnumnere numer oosons vne numer o maceosons o maceosons, osons no co eoa numer o mace an msmace e resu unoneasneeerc wennaowe o o s ceomuenarson.e.,e wnow sze, an mu n wnow o comarsoneweenwo seuencesseceneene. ennreesse oerwsencae,ewo seuences.en oromoo n oe sorer oe e w reseco suc seuencesseneeren ase erceneas, ae oer a amnnon ace r sesuesne canae seuencea areenca wenown ercenomoo, an noe counesncees annroucnas, necessar,o acevee maxmum en.-ermna,-ermna, orrnner anna e conservave susuons asar oe seuence seuence sa xensons,eeons, ornseronsnoeee “ noe consrue as aecnomoo. no amno acs ceanne m.uaon” occurs wen aon muaonrouces a snonmous coon w nvoernnoer:oecsseo.: - - exosureo a “ns e eno a vn. raonnemouesn mau aaeonn,”us a ra muear roensuos arom s soenuaennecoeusao occcurs wou ananace wn a ce,ncun err emca reacons “are nucec ac”s a nourcse oc recaon. aare canear o a cromosome, esome a,c or vneco wc aransosonsoensere.uc n “neraonare seuence” or “are sor. s a euence” or “are se”or aransosaserans soesa osre. seouren ecxen aare moecueno wc aransoson canensere a an-ase mecansma,m wec, ac-eransosasesransoseerransosons usn a cu-no a moecue.ea rreseu ssenuenucecsaon oerasearare seuence onnseron eransoson usoun on eac se o annerae ac- rosome “reae. nerm” “era snsaon” reersoerocess wc aoees snesze a “ransosaese”uensce a oo aonuceoe. ransosonrom aonoronuceoe,eor eexamae c aaazese excson o a corresonn noneraon-ecene suseuenneraon oaesmran,s anosonronvneransosases ransosase mae a ac-eransosase.on-mn, suoa aearreans nuocseasce ac. scosen..aenos.,, an,,, eac o s are reerencens enre. wcsncororaeeren rom oneoenerm “ransoson” reersoe acon o aransosasen excsn aransoson onuceoe, ourcneoo a seec aonnoennunceeoraen., eerno aeren sene same nvoernnoer:oecsseo.: - - on e ee arm. “ransoson” means aonuceoea cane excserom ars onuucceeooee,, oorrnnos aan sceec,on aaosmnuc, aenoeeneraeno a seconosonne same o a ce, e acon o a corresonn,raonrs-ancsanncer,anseoseansoem.c or exracromosoma arsransoson en an a seco ransoson comrses reco nransoson en, wc areonuceoe seuences onnuczeeoe a sneureanncseoseeween areanswosase.ransoson usuaurer comrses ars seuencesransose aon w ewoorarnanssoososonn e enns,s sucea acoen orseorannusceoe auraransosonsreuen comrse encon osase. ransosons aransosasea acs oneransoson. onuceoe a sse cueanmceeaesrernans aroesa “sene vrcuransosons,” comrsn aeerooous ens. e osuxaosoneweenworansoson or recononeerm a “nranrasnossoons enon” mea an csorree cs-acn nuceoe seuencesa are sucen ens o ac sonnransosase.or exame,ransoson reeasnewo-raensraonssono esonnss c aorem rerseerec ormerec reeas sucae resecve asermnanvere reeas or “s.”verrsaen csomosonem enen ms oa e oarc oer.ese are reerreo seuencer eonxmaoe aromoes ma noncue an aonaransoson or aumensransoson.on-mn, suae “ es ar :.”secnoss euc srseos o reeeern rswo. se s.e avaere aan renenoes ros. m an, er,o coenn a esn x,or re w, ec,.roves an examencuese secc , as we as aec renceo aene orroen aeas,, or en ereo.xames o, s aeceesc, a ann snecucees v vararananss canereeoouanvnn nvorneoceo.: - - eenwosr:sreoss a en a o.eraaases. n one as asm-ossems asmneenen oec a,n aoascm se-ecoson s asnemasrovea aowsor manenance oe orn o recaon an seecae marer.n ones a as ceocm,areave smaasmacone.e., asm mumerormaon an exaneasm seuence caosmma-os s.senm o sneow as reuce asm-os ssem cane causeorouc sec,e an ex e moreasm or moreroen wennuce w aen,o orenoous. aenucn aen, or va auonucon uon consumon o a reressn roar , wc areen rsearo am uocecomnanoses uceoes coons a ora reo rroan un oscsre oer reo- e,oe cre o cn eoemor smeecnacr amonrmos ac ons mnes aseanreer roen. one oe nuceoess muae, suc as r e m seuence a aon muaon,e muaon ma aec on muaon ma comnr usemaee reaceemroenen o.everaes oon muaons exs.rs, a canen one m nuceoe, an umae one a caneasne oonre an aomneor, w acc maeao a muaon aecs a sneasear.eon mu .uc aon m coon coes aon ma consue a sen muaone muaone mo crooen s caomees amorn ao acere.eon muaon ma consue a mssense a conservave muaon or a non-conservave mn amno ac, wc ma,nurn, ma consue nonsense muaone m coonecomes au saoo cno.onn. aoenren muaon ma consue as arames muaon, wc manvoveenseron o aase oreeee oone one ac masuearoomn nvoernneoceo.: - - reesu oror: nn maess e a se s.eueunecnec,e a.ranm aeso mnuoa eone wre aeson ca or crean an aonaasene sacroun, aosa mae use nee reanrame oe m. n one asec,eos comrses aroaroc recoumnnaneaossm ce-o.s sns oenme assreocve. roaroc recomnanos ce comrses.o.n on ,e os ce ma exress e asec,eroaroc recomnan areneaas a uneeerroonoeo ausraomensuceo muea coomrsn an oen reanrame comrsn remaure so coonenresenownsreamnen r,eaenrarmamese.n m onuea asone resunn a a uneroeserames muaonse w-ere c,erene erenea uneroeserames m ne :.n one asec, asec,erames muaon uaonser- ene :.n one en comrsesenseron o an aennean “”noer- come.rsnes on ane asencse,roenrnemoauree soo coons .n one asec,erames muaon reaveo : oer semeneweenosons an numere muae orra - ene.n one asec,eu seuence oe e muae omrersameseer-r-ene wene coemrnsseesroen comnserrseos :.n one asec, oonesrervoanve muaon comrsn a n an, oona, aeas one c sen muaon anor a mssense muaon resunn a oen reann m ounrea am ason ee ccn o,mne r a resu sno anno re excreassonnneaoererorooeonu.sonuceoe comrsn an roaroc reneaas unerone arames muaon,e comrsn a renc oomenn reaannosr caeme ma exresse same or aereneerooousonuceoe uen recaonnaorroe conm orr csonnse arv wave vaeranreneere oor.r enea coesor a nvoernnoer:oeceo.: - - ce.n onenss a osn e ee a c a,s.erco,areo mcu raeceom ornraamnesose crenesneraenoeenome oe e muae orram eenomesecenne recene, an roaroc recomneasneos cer.enne onsen aseerca,een mouaee recene oeenome oe orne on aasm anuncon as a recaonacoror, e.., a onrrame osrne o rerecnaeo mnae one asec,e muaeec oarornna . n orroens roen.n ameserene coesor a recaonnaor roen com onres ansec,e rec:ao onr an vaarorroen comrses aruncae recaonnaor aon, oreeon. varanereo coamnrsenreo a c coomnserrsvn a conservave susuon, eeonns conexs onen wceroen reanssuncaonvae su ass a rueonc,a aonnon,a oorr asroeesncraes cearean.e o aecr svuaane a conona orn o recaon as exresse aasm comrsn,or exame, :runc oare recao:n.nan oerro vaernasn msa onec reuecaosenaorroenncueose w co-u muaonsavne on roen : w one or a comnao asc seuence oe w e , , , , , anoreeonns o muaons seecerom , , n one asec,ransaon oe m o amno acs- or-. mancueransaona sae.us, wuea meo osr orameesransecrr wenee acrraosseoracoersrameennn we sar coon o nsae accornruncae re eene ae’ en, resunn aremaureransaencea neoxnnramaeo,r surcoen,ae reouso-mee ma “s”orwar aeo semen an weeraee w-eroen,eroen cone roercaeonrnaoerne,r oore ann oserro vaurcaen, nvoerneoceo.: - - escnroer:ess ee a am.no ac susuons anoreeonsse aove. soa resnu sosmne e asecs,ere ma exs annerna sar coonownsream oeremaure exresson ooxe mreassnonro o aroen wose-ermnuss noransae.n suc asecs, , or, as we aseuuecn o reeracmaeosnnea.o.,rroroeenns ceo.m.rsn, e.., : conservave varansereo anor aroenrom annerna sar co,on’ oe:ram,es, or mae useuo suor recaonrome or-cona , some asecs, sucro nnasmsescreere.n ma encoeeruncaeen rse macaeon exnresserom searaes.n one suc asec, one -ermnaoron oeroen.sraaonrsaroonean, s an annose ar meaa encoeeu-en or some vruses, anas asoeen ure oene exressonn oenmonsraeo occurnacera. n one asec,eeerooonaousssenaenes essena onuceoeurer comrses a conona coeneeanree cosm usresuno arenroaucnn muaon.aes aae o conona essena. asmacone, werene. coeeneaarnes m cuaauseon mseoaro comc aauxoro we ama omarow w aona suemenaon wen cu sa sow onsrucon oscer uren mea.eeaa, e a., coecon.osoc.a co:-n-rame, sne-enenocou muans:eeo : .enn ..o:. ms .u e. encon-aceucosam-nee-cr-euoxsa,e va unrereannosoer.encaon oemene c :-.o:. ... rasenscerca co.acero. .u,un,u,uan,u,-,.ao.. :caon o as a: nvoernnoeroceo.: - - nove se:ecesosn e m a. . ournaarero meaeene exresson.one. e :e .o: ,on,un.on,e.n,w.a,:o. novea:a .m,eon,m onm.one. :e .o:. ounce-eaos-vecor ssemase : : , eac o wrcna.sonnec.ororae.u reerear. enre.ese auxoroes cane overcome exre ncens er sson o a suressor asescre uee rmnne. o uornoe mcaeaooesm aeenunree or aueneomsenne nvorneoceo.: - - aenor:ess e a m m. osn-no-aeccses m-ea-oucsomsa amnne ceo wsaness m ac-eascoe-,-ucaomsamnonem oerc o-er ean muaonsor coma w am . co uu ucn . co aasmacone, weren muaonsne ucose.eneeearuesc coause meaoc auxoroes wenrownn mnma mea w. ovo. ec su:ress-on. uno:er.ns memaooecv emvosmva.arc , e a.o , wcsncororae reerencee .u e. : ov renns enre.ese auxoroes cane meercuomme exresson o a suressor asescreeren an canerownn rc en suc as ro wou aona suemenaon. aremo e-anceerouucamae snase nvoernnoer:oecsseo.: - - c c cassr e a c. aarenam-on-su-naoesransancaerse snonaeasueaa rreeu sauourns nvoernnoer:oecsse eo.: - - n na a. aa amu-ansnooarneaaoxee ocxoosrassemoraeo ssnraanssee sruasuenecaroxan nvoernnoer:oecsse e ao..: - - ecaon ounae-oenrvroe oasemxsre rsesuonre oerene recaonacoransoerons serouc oerene as a recaoneos ce. ne amma orn o recaon annaesnsee cen a concenraoen w-eeneenr maennenerr.ouc,e,oormosrmo a monomer an amer monomersaveeen sownoncreaseeasm co numer, ansmenrseocreaes oer’.saerroons, muanroensaveeenene an canave a sncan us eseenoes areounarea mac onasm co numer. n more oress ecen ann varousr cooneuruaeoonsemornooemnesr’ o armeroormmers oro or’serons.n aon,eroen auoreuae oe amma seuencesns na ss own exresson nno oeraor or w e,smovreanromoro coern.roonnseuaesnm, c coonro nu o exresson oerene,e muan ce.verexresson oerene caneoxcoremroerarnoec rreocoamronacn recooms cnanos canace an unwane meaocuren oneroa e anor numersoo urn exone roc recomnanoseasm co co numer a a reaveow vanueaurronw e.arn sormowe ca oses,e mroae avanaeouso manan cuure, anenoncrease aroc recomnanos ce as e co numer soreorearvesne cuureosoaee emasm. as aowr cooac a nuomwser,enroe aarsoencce re oco amnanos ceso recae rous w co numer oncee cuures sucenenseo a moewaoor acuunraenn, re ancaeonn oave aeuarsn woue neeo aso recaeeroaroc recomnano sm os en, aroaroc recomnanos ce, somse ces. sran or aos sran, mae consruce accornoe meosmesecsr reeerreereonoer meann aasn a nvoernneoceo.: - - ourn raereor e c: sosners eos e o a. romoe eexr.resosnorno o o eexrresene reacne naveromoer w annuce emrca omz son oerene maereoree manuaeo cases, mae useeuroou ecsaons o aeowa esxmresseonen an one cuurn conons.n some eneoware en oe cuurnero,n wc cnase,e annnncruecaseeero emxoreesrs sosne oer e emoe, wnucer ae a an aroraemeonuc m wou mae useuoecrea e exresson.n oer cases, a reresseromoer osre ex wressroanw oa oenurceonne,, e w.c coue accomse eerrounucer. ., vauon oe cuure an an aencue an erxoomoeneoru ssse cmonsroaa coeuromeo emer soseemo accomssunae exresson ma os ce, suc as, unconaneroaroc recomnan ures anu us nno amreoroa,er roemroesesrosr consruce accornoe arcecure sownn s: an-, oer an acvaorroense..,oseearn r aor seuencese..,oseearn s:-o conro aacomoroemrorevrn ss exemrse,ssoen- oon aenreen-oee. s.,s :n varaons o -nucenuce ssemse.., ara :em,s r,a cmumnoasee--nnuuccee s ssseemmss,e.-aranose- :,osae-sensve ssemse..,o :, or. m,aae romoer: nuce or : nuce.n aon, sanarnucenuces maaeea umseao erxomreosesrsee onre suc ase amne oraure s: an-,emeraure romoersa are u-reuae saLrvaoRn, sress, or enr-sennos svaeo renauaorroen, c, secc reuaorroen, anromoersa are u-reuae sarvaorn, sraesses, or a en reruonno- nvorneoceo.: - - sae ooonno ,arr:es oas ,se e a. o,oeae ruecunea roesroroens,ncun aernave smaacorso,o, r nsve reuaorroen , ane caaoe acvaor een. , anorn resonseo sma moecue aarmones, suc as c a or :. suaeermnaoror suc an exresson cassee maeeermnaor auonuconn, oonre e axsamece,,ne resnouncseeo a co cnuroure o-ro exucreess nounr oenmearoene mancue eow exresson orenesneoe aunance o a nurenresennn.ne mse scaen aaroe,ne annnnncre oaseen cuurnrocessasraua consume e ces, conseuen resun se o con erxor oessoenerorooms nuren-sensveromoer oncee nurenseee. rouc suca no monorn oou cseeneens exreosrsonm enaeseroucon o avenene reure.s aroac maearcuar useunenroeuc aon oona ore ex aomenousnucers uo-nuconase on nurenmaon maearcuar useune conoeuxn os oromasomers. a are ason su oneec asoec ca, aaoose s rreanresosron.ma wunae an auonuc rrene mae creae rovn a ramnose-nuce exresson cassee exererssvonn o aer _oern.romoore erxame, a_r:a,oroowmeoererene:se mae cone’ oe nave , orenes conorroensen .., an o s:,, avneasc seu e s:,,,,, or co-u muaons muaons seeceroemnce o,e w,ero,en, ,: w, one, o arn a coormeneaoonns o o nvorneoceo.: - - a omennoo s ar cc: asssess e ees- a. or-:,. ann o,n oe asoneac, as exerrmessnaor :.n exame e recocusn on consruc maenrouceno ar*. a sran conann,or exame,eenec mocaon oe arene manan aaassam ex wress coon o numeerraen wen exeressose sraansmes-crornee s mae suceno ereea w suorrowne asence o exoenous arnne.ecurocoremssoeren ceso a sos mae mae accorno sanar meos. su o eecrooraenoe ces an se aemaasm mae n one asec, aasme mcae oenro mucenma c mueuran weou smua-emene arnne. an arorae meum suemen conann srann eer e wucose.asme omzaon exermen ma eeoonrm oeucoose,e anevees sarn concenraon oucose an ramnose, suca romoer corresonoaon o caaoe reresson, annucon oe ramnose-resonsve conseuenncreasena osmma co ce numens, resunnnuce exresson orene an asmer cuure, aseermne e sranera cre. menoss wae.,.,ereaes amoun oure measuremen, mae aceve. cuure an nanoro n one asec, aasm rn oasmescaon one asec,easm comrses ana o mrane o us reeucanoneasm-os ssemsrove.n recaonnaorroen exresse eos sran.n onea assec ca,aee or on ac ov raon e -erve.n one asec,e orn o recaon seuence ma comrse,or exameec,a anon os nvoernnoer:oeceo.: - - : orss:s e- a. or-.:n o.ne asec,e orn o recaon comrses eroaron ocne re acsomec,naneaossm cesue erec xerra e csoe semrarsee ars seecae marer.n one asec, were conona s aeerooousonuceoe comrsn a comrse ee ers:sena a sreunceur caomernsen s aean muaon,e seecae marer ma comrsn an ancoon caae oarenue wnce oe a c sounreosnsoar esse,e suressor seuenceeneo comemene co naene or au roa nona essenaene.n one asec, weree a conroocna recom esnanos ce exresses aeerooousonuceoeaoes no comrse comrse an anosenc raesseannece co camssrsn aean muaon,e seecae marer ma an o n one ass:ec-,.en oennee a seeuee.nce oe suressor mancue,or exame, s: an.n one asecs,ec,eermnaorrso mmaoernsc muae,nocue,or exame, an o s: an. r exame, an o n one aeseerco,ooeusaosmnucueoesnconroens oneres recomnanroen oneres an an exrreerss coonm crses a nuceoe seuence conor a roen oneresn aos ce.n one asec,e recoamsseneanor exresson oe recomnan roen.n oer asecs,e recomnanroen oneres mar cooemnr osen aeress aeraeuc orn vroranscron sness, vra vecor sness suc as vraraacnsaonsaseesuc asranser vecors,roensrouce recomnanor acaonsnoo, cosmecs,noesu aens, nvorneoceo.: - - arorecn uuor cu: ornaess .ee e a a.n nurona suemenaon,nusra cemcas, an consumeroo coman one asec,easm ma comrse anncoma a rnaveoeoso srsa onsecaon esx cora resrcsasm e rurecss arameserene asescreeren. aon o aroarocecomnanoseorae n on nenance o aasm oeasm.eo ars eexcam,ee, wroearroc recomnanos ce mae enneereor es canes maensue: an aeeec reroaacremoecn recomnanos ces. co,woenec muanenea coners cononauncon onn oe areosesncne oecene suc as ar w a exresson o aenea coesor a recaonnaorroen, e.., a suressors:,, an,, w, o orn ceo or- au com munaaoonns oav mnuaoenasc seuence oe w eroen : , anoreeons o am s seecerom , , , , , , s no acs- or-.o accomseseenome es, eannomare eenonm,e r enecnues mae emoe.sncues,or exame,as-enae srae, suc aseacemen va casscaomooous recomnaon, an an aec reacemen erenns escre aseno ananner, , wcsncororae reerence execuen en an onr aee e as. ce rce,a acemrsen m soraecaonn ooeenome oe. coos sran mae : ore mu rero exresser- ene comrsn n one aseacn,reene corammresn :. eenome : or ma cnon usaenoenoroowucne ese rmeencsa:on suor eemenno nvoernnoeroceo.: - - .:ess ew a eo. e reneoons oe eomeo,o eac, eacoen ncuaceooeesosneennom.enansn eer se o ene mae reace we aroraere case,e rec e muanre ne exresson cassees conann encan nee.., :, aneannomoo arms mae exresson c saessueenecenoo aeern’a anve s’ reeons o recneos.neraon os . reon o conannerss exress conemae. con ene exresson casseeromoer an an . ro sero meoc e eroem ,ne sr mn ucasr, s urearr,uc eou nrra e, e axnaemne, ere a m,n aerm naoarnanmao ancr en. ressanc see esx rerceossnonze cassee eecro as reeccom,en e n one asnee.ar ramen mae eecrooraeno aranose-nuce ancaonmnere,en ce.sn exerreasnsns maameasoe recomnaseromasm aseno eanamcn.ne ae anor ressan coones on w r raon oe snec mae vere usnrmersanne rowon osn oaromenocon-ceoscnranen aove.aeosss o asm mae vere aaureo exressne recomnase an an am.c senco renssaanscme m,arer, s maenenrouce aneanamcn ressance marer mause remove e recomeencaese. onearenc ann, roeme e rexsuresnsonasmsaveemeraure-sensve orns o recaon an mae cure an remova o sreanneraroew r aessean nonermssveemeraure, °.oss ooasms arencn an ancaenamenecn maneera veereene.n asesrenocne o oerowre onne norneoceo.: - - e rexvernecso asro: onn. ceassss ee ae. mae vere va an manenance o aasm we orn o cononan e oxnere assseec a,ro r*eeancee, aeneoasrnecraenmee,n e. m.,a a we snees azre.enne su w a asec,enear ramen maaveeoow c an . wo reons oom n seuence eemens :: eeneoe eeo, eoa, eacencaoeosenomeann eer se o . c o nuceoesnen, co reononn oace o a coneannnaannee a cr* aee, wcns case maave a so we as an aere coon usae’ oooenn aro resuceue so o coeonoee can, as . a seecon marer,ns case aanamc , romoe n ressance cassee w arorae reconzre anermnaoror exresson osene,ane ses n on e recomnase. oowe, usnee as mec,e sameroceureescre aoveorereneneraon mae mae vere aoauree o sraen r exressnerene ase sarnos.ec reacemen u no arnn esun sranorow on mea suemene wucose auxorocrse ow se.co on ee m cuouansn csoeuaaer seu anor oee aenerosrsaenaes an, ane escreneaes,n e mea suemenaon as eooru e asence o a suae suressor . n one asec, aca moen ooseraosvmenore muroarcoaconecomnanose comrsn: cuvaneroaroc recomnanos ce o, weereansme,reo mareooc nvoernnoer:oecsse eo.: - - recaomnuncaensnoes a rr cue. coonm orsees ara cnosnona essenaene comrsn aean muaon e muae ce nroucneaasomn one conona essenaene an auxoron e suressor oeasm comrosese anro aaroc recomnanos ce,n wc conona essenaene ans secco an amno acnc coon caae oarn w e e conona essenaeneneroaroc recomaae o resorneransaon o asmrome ce cuure,e seeconen carre ounannos ce an seecne oreroaroc recomnanos ce’srow an cuv a meumree o a nuren va eoorroucn ae ane seece ces. n anoer as comnanroen oneres e meo comr ec, a meosroveorroucn a recomnanroen oneres, aroenroucosnno:s mu ne exressonasm asescreeren cuvan asmaseenransorm ceeweere ornaos ce or a seconos ceno wce oneres an ur n a cuure meum aown exresson oe recomnanroen eroenroucone recomnanroen oneresrome cuure meum orrom rovenna onee asn .eco,s ceer.oenrouconos ma comrse an one oeaceraoss anmeroaceraceaeeenussea neeucmeosnae nvoernnoer:oecsse e ao..: - - ssseeeuuuooommmooonnnaaaaaaccceeeaaaeeessseeeuuuooommmooonnnaaasss m s suernneoracena * s n one as eoorrearn aoeuaroce sruon.e ececen,ce oansmerasoanv,eme srzoev oe caeaeces o euaroe ceeneraon annerae,e numer o coes oeeerooous seeroueoncoeusa are seneuernaceeano ca enace nvoerneoceo.: - - eenonmo eeer r,: o aness ooue e a. se oneono amracnsocos wonereneraon occurs maeurermrove acn comrsnnra onnseec asnec,e a m-ceeo ws. aroavsme asorerearn a moe-ce,e meos comrse o ar screeren.n one asec,easm ens aneransoasnosno esnons. aren s onae asec, oneenec maeraane eransoson onanecaraoensnoroemaenome oe-ce. aconesecama useas.mor s exsaemm me,aee umseana a casaessmns wc a sanarasm esraeo avoe neeor sm ssem mae use weres reuaoruroses.ema anasmoc s seseecmo mna aneo reuceacera seuence conenor conro oeasm co numerneos srano a uvsoe m weeareocs oresorxaceouraveeunae eosrowase wen on a mnma co numers nurn mae use were a smaasm neee.emaasm ssemransormaon orransecon ecencacueoon eeas sez oesaesreoreurose oncreasn numer anor ma sm enr orncreasenasm co avanaeouses noms. wansreaasmm a scsem ma asoe use were reuceasm mumers are r aons.eucnasm mumerssmoranecauses meuucmesercacsmonca semzee.reucnasm mumerss aso mecansca vauaeecause mumerss aso mecacnsnscearon vsau araeeessec saaean snenserons.eucnasm oasa are unsae, an wen mumeursse a vreecoorrsmeor ann v rreoranscron encoenruncaon oeen oea.umers aso aear more resssoavne,onse warza ooenn, w resuc nvoernneoceo.: - - ma coonr:reusse e a. eo rmeaaroasumranscrsn m rearaons. o euaroc cesorroen sroseumc mona, vrea us veecnoreroucon oasmsorransecon eneera, vaccneeveomen,eroucon oraosmucsono,r sae a cecaneonesv,eomen, sness, aneroucon oroen or meaoes ncunransorme wa naceraermenaonrom srans aconenn co omnen aassm oecn,s. we oemraeneacc eeone mae usenace o a sanarasm useneenec enneern o euaroc cemse,n ass woer aes vurose o crean vra vecors oen eevan vra vecoresncueenvra vecors, recomancacnn aeeenvoeomen anroucon. amma rerovra vecors, aenovra vecors,eres smex vra veco-assocae vra vecors, vaccna vra vecors, annuenza vra vecors.n vra vecorrours,acuovrus vecors, w e vrussve amon severaa con,eenome oe vraacan smsnosnc seuence moues sucae ucasms a creom caoenen “sac area nononaesrane e seuences reureor vronacan. asm,ca caee “ranseramsms.,”” weca ccararesn see muoencees a vrencuen anoer enneereenec caroa weacaen raenome we ca usen conu oe vra vecor.eranserasms cesorouce vrus,ncuon m wa aseoaec uaseno sansmsrann a coransecon o vraacan revouseeneneca moe sec aacan ceneaas v o exresse vraacanenes.erocess o vra aneco cer m caunuuarec.urn curren reuresareuanes oasmsorare scaeransecon nvoernnoer:oecseo.: - - wc are anse e rn ex a ca.maeoo o ao vreane vreaceor crmoeurcc aonnreonce rsesceenoerr-aesenvra vecors s, acaons.o mae s,reeacanasms, eac encoenarne vra cerso, amon oer ore assem anuncon oe vraarces, are neee.ers carreseeans- reoure enconea anaooroens, wc are u ene roens an maerocessenoo srucura revene, w enczm enesnvoven vra assem anransucon.eevasm carrese unsce anaraco sesc ae m vura-unacoona m reuaorroenaacaese exor o crucaoreroucon o vraarces.ero mmu-eu nnucceounsaoe coasm, wcs arcaesn vra acan.enveoeasm encoese re vuraa eonrveoroeenr aso wc enaesnn oe vronsoare ce e oen, veco xraceuarroens,eermnne vra enveroreorsomee rane oos cese vrus cannec. common useeerooous aowseenvnrasarc-esoernveecrome vescuar somas vruscoroen, wc carrese moe vraenome a wroae r eaxnrees os cees.eranserecorasm a weacaeno vrons aneverenoeaorn ceo cnesrsu.crevneenes oneres cs-acn eemensoreranscron anneraon oe consruc conanse necessars cusomzae,eenn oneene oneresoe exressenenaroee ceo.senome an

[0002] nvoernnoer:oecsse e ao..: - - ranamexamoe:maosranonsru ncaaon ene naarecoenuence nanerae . arn sar.n co. s croan sran,-o-:o,esnaeeren as , wasurcerean enaoe..ran aseenoe:. co–omsr–a mse–rcomm+ nvoernnoer:oecsse e ao..: - - n.seron. o cor- sranenen sra an rec::r- eeon o rec w concoman recomnaoonn,s anrucoenns oere sen euenmca: eos sran:s w maso accnomese. co vaenoomooous nae.omooous recomnaon messe esa meosor ene. coenome are we- nrousce anncue suc meos as casscaomooous recomnaonase uon unwaneon o ar roessanceene on nonrecanasm w suseuen remova o wnear mmeaeeenome emon a counerseecae marer, aec reacemen enzme meaearee ceavaeea wm aan-e recomnase, or asor anoeras casseenoraens recocus an o v waere a o cmorno osorrua oce ue rear s recomn csoenrs nanruc on a. .r-er coennser euxcroesson vnercaon oasmoss asence orow on anoc-conann aes, conrr aenc raensroarmone o weesre seuence was vere va , an eac sran’s a oe e --eaerarnnesasmas w secnomcn ressance cassees wasese. comrse “acones” comrsn a ssmecsn aormecsnenae.ac oeesasms romoer_ oe orn o re recsasoannce cra_ssee a.nace semenrome an n asm wasess asmuceornes oneenca.on eaercerves acone: : nvoernnoer:oecseo.: - - meos.ers sa e ens a. wo srans a ann we were mae eecrocomeen accorno sanar e orasm uress reransorme wasm e - ecnomcn, anncuae overnor a, °.aee orenaosearenara saraesn w m ve rseo secnomcn-ressanr oes no ee a ansormans w eerasm, srans anneraon oenr a-unaennce o secnomcn-ressanransormans,ncanae e -e ane coansruc wasuncona an suore recaon oasm .. co sm uressor . : sran uane arenee coono a so coonn sran smaroueanorno oucoen oee r ceoco:n:or- aen seoee cooonnnsne srroann usne wase seroureme : sownnae.ecauseenrouce c nce sooransaon oon w resun aremaures execeoave o suecuen-ernee arroen,oena muans were seece on, wc osnecawa.oss o anronnce reossunanccona comemensesonne arnne aes. ona,ransormansarow one wasuc coosnermaer wa-cn on arorae ucosear wou-rnne, are canaesor successu muaenresnsne,u no on coonsoe so coon. anoc sensve, oee sran -arnne auxoros were save as m-arnra- nnes . anncua ane overn we are sr °ea.eonmaree oonear wno m oonero w m-a srnnen ceo aonn o ro eaec o svrearnn was anocua °e. nvoernnoer:oeceo.: - - nramnas-reess e a. an an were ma weere ee scorroecom ase sreans an , resecve. eecrocomeen srans werer n accorno sanar meos, ane uressor , aon wa ansraonrmseorm waona csoms e -e an wereaeo araes w mecnrnoomaccn .eransorme sransncuae overn a °.e exermena resus n an ucosearaes an ae rarnansorma xono -oon are sownnae. ecornmomaocnn-oonucoosremaon easm e -eeeransormans on ar asmmecnomucn,resusor noeeeranrsanosrmoramnas on ucosear.oro srans, e ure ns on ucosear,ncanaransane arssor s cae onsern-enaanne ae so coon wen .. co sran m ovoen rasneeoou as munucaoonnarroen. r- ene n resunn aruncaon oe rcasm,ons a vee esron core: wauncona oema vecor ssem w aarer an moreeaure- cassee, an an aonas c soensmreunce o seau ceonncea snueceameanaa sczeo wnaes, an. amcasmn ressance nvoernnoeoceo.: - - uresrs: eoress e a. -e, u:ress.oreen,e a annmen ouerenesnasms ss sowneow: essor encone : e -e: :uressor : :uressor : eecrooraeownevoer,s anasm seecnea onaeucooseear.osveransormans wen severames, an evenua ecrooraon was reeae asm.easm wasso aa ceoornom wass s sereacne anaou snuooreave neo r meuacaonosn, oees aeenoe resuerom aenomc muaonneos s suesn easm ourow on, resunn ran.s sran was cure oransecon o eu aaroaocn ce mnneso weneraase s m ara vn seceoarr w. a smaacera ornor usen comrsnuressor n aea-n™ransoson,veasms eac. n : an aea-n™ransoson were consruce, a acure aen mu,a anonr eannasornmenoo r seracnaseese anasms.oee cson suraranons an nvoerneoceo.: - - comnnoar:ones os e ee am.ens oeveasms are sownnae. anasmeauresonuraonac:one : orromoer : erm: e resuscoon counerae are sownnae. nvoernnoer:oecsse e ao..: - - oansm. ecnomcranns woerrema onns oaserrveew onn s mrannma mea. woo cuon arornne or on mea w ransormaon ecencesewe orosseerencesn suse oransormaons was reeeanee, cuom aseoennc cuerearaonsorewo srans, a smaer arencn,oesor r easm , conerrn ressanceo asm eave ecences.e resus are sownnae. n nvoernnoer:oecsse eo.: - - ar w uere aams. coare snrcansn,vnec samnar naumeerwso o srans arer sansormans on oone oasm , on sran ves rseomaransormanms oanr w reusceco uae oonear um secnomcn ose aar an onuraon ano.nuraon resuen s eesescrvoem overn cuures o marasm r ransorme rownn mnma mea an , o.en wease unnoamecoe seurmonr recaon oonuraon . maasms,enomc rom sranasnn ana ena wesas esocen recaon oe scercenceexenomc urcaon,xoranoaoree s uesnuen acermo umnae.eenome o eac sran was seuence.er- enes reresn, an :.e reevan reon oeenomc o ene reaveoe sam e srans sowneow, -:,e re,on oeenomc oe sran: - enen srans: se,ue,nceu vaarvaeoonca coorrnesso unnseron an aennean “”nseronner- muaon suca aremaure so coon, ae.enseron consues arames reanrame.eu seuence oe muacze,sresenne resunownsream :“r*ene”.e seuence enn aeer- so coenoens o rerurenscene :.e muaer- ene enc aons sown a :: oesor aruncaeroen comrsn nvoernnoer:oecsse e ao..: - - seuence comesae exer amcreonsss v saroowusa * asm ,earnr*ene,ossessesroaer u-en r- e s comareosaren sran, , ws conona orurnexnaem. o re scoawes: on a wnra en oxs saemeco unen o conmaa-ces na w asmroa cromma ocr cso onns s:rucs ce re auene encon ananroen seccore orn o recaon aneerooousoe res a recaon aene enconor a suressor or an amer coon an arroarocos ce aene oneres anenesor reuans exresson, ansoson comrsn ens.ure sows a sanaracone con eransosonane ransoson smar sze co ro.ure sows a sanaracone, aove.eonreero a s. -nec™ca,-eranmsaosoasms comrseonuraons- asescre onroasm or “onro,” as sno wwansne saumeoreemaasms ane sanar orn o recaon, as we as aanamcrne re.e asm conanee or “onro,” as sownnure was a smarssance marer.onroasm a smaerransoson ane sanar vec sze asema vecorsu conane rs roun o exermens waosr oracanoznee as se aonranna.e. nvoernnoer:oecsse e ao..: - - mavasm acone. ra mnsosase een coae wwo v aans- o a mn aonn- ces m eaocnoc woenrea a acnvaoeeso eraca asn amoasaa °.e meum wasonza-vo-, conannea-nacvae rours a suemene w uamax m-aan--uamneeeuman serumvv, acvae ces were coune usn aecmanouer-e anna- m.e c ceensruera mo.n, waes cee o mncexunre ee wcrooraonuer, an resusenene samzeer,u coere acex8concenraon conssen across samase ssu.aecme sname we was eec-ro morae, a anoweasmo recover a a aen a sne we o a weae w , an or anass, ae se res meum. was remove,ee cemeons, a same o eac cuure er same recsonouneaes, resouseenen uer . , suemene w o ow comeer.os-a en, an anaze usn aonoecnoo anaca oscences. cusonaa anass waserorme usnowo soware secon roun o exermens was oranze as seornae. nvoernnoer:oecsse e ao..: - - maevne acone . ra mnsosase as usn ancmvae cesurerom aeuoa an userec were acv e mmaunnuoacuureru’msa aeor accorno nnsrucons.e meume wacsvoanozra-vo-, coecnooesnacvaeuman serumvv, suemene w uamax m-anannea- ee, an an--uamne coece ceen arc- uvaaem on, w cae. sse w oenrcee cnounen uos cn aecmanouer-enazer, eecrooraonuer.e c acum or manesum, an resusenen a an a coanceennra ao snn coens wseen o ae acro mssx w saumre weass. suace smamenee w was eecro-o mrae, a anoweasom recover, or anass, a eae wres meum. was remove, e secemeons, a same o eac cuure er same recsonouneeeaes, resouseenen uer . , suemene w o ow comeer.os-acen, an anaze usn aonoecnoo anaca oscences.e resus arues soownnaau arneass-s wa ass aveerroarmese o usmna aonwoon sorowarroeus. nvoernnoer:oecsseo.: - - cesn eac cau e ere a. or, saoswes ueono coaunve cee.oave ceencaese cacuae ure sowseercneas an cuure voume. ercenae o cesn eacouaon exenuorescence.ercenuorescencencaese , wca ressneuorescen reoreraa exceor e cacuae cuere. sn e sacow csaeeoruoraesc weenrevvee c aenueor.esceuno,resacsenve ceencaes anercenuorescence e uonoave cee us,euoremscaene,veo. ceeeoa—avree cee aneercenae o osve ces—an eer o erorose cesransece w maanor acevn aer ceoanerrora snamseces.sncaesaemaasm conerse avanae o ures ane c seowean reevan conros. oee sow aa oransecon o ces w maasms os-eecrooranonn.ureour remaave voe conros sownnures anor,ours a our ecors sowencreaseercenransecon vs. or sownnurus. ere s reoawsveoea coan orora snoswenconn our cees, woumrsaos-aesmecrs oee ourma vecors sowencrea ooraon.. xame seercenransecon vs. a ours. an:aorns mreucaonor o a. comarowos anaasmowsroasumceecon oncea rc mea.n some cases, rc mea maereerre over mnomnan mcueeaso mnma an ca suorsaser an more rous cerow anereore ma resunnreaa rerc masema nvoerneoceo.: - - men a.o enr: neees a as e vesen ac. ae oec ceon.sann a seecono,n seerecssounre ororavnseorme m snramnas o mne rca, measma a raerenon mae avacnaeous over seecon on mnma aes uc on r meaaesecause coones wrow moren rc meao s cuo m cnesrua. ac s aeemaos srana cane seece on rc meaaes anerown a waserveromecnoownnorasm s reraenn aonnsa manane, an. co sran was cosen wc encoeseroen seuenceor asarae semae anen-erameroeeneasoen oe asene, resunamnomec ac auxoron r .eeonsn as meaocawar c mea sncee enzmes reurene sanar rc mea suocuc asn.o,r a c cornscruac coomonen oeacera ce wa noresenn ucena.o. w n osos sran,e-nransosase w exre as useonser aonuceoe consruc : :,ssron c*asseenee oseorrae m counaneece asoe aner womo aenmer so coon ae , an coramenco ress r an aermnaor s:,,ransosonr ance : ane nransoson ens.e eecrooraeanmoen ee wcarsocome-aemne,urase, ac mcoxe w -nransosase, an anae sransor successuransosonnseron werer consoe manuacurer’srooco. . mo um coramenco enrom coones seece on usnr , aneresence oenseron was vere w .es seccoen ens.ne suc coon was seece as aesos anesnae oemaeesmaasmos woansesureaoonrs aransoo srumaoorn s oee eceocnro ancom maenenena cneces nvorneoceo.: - - ane sneo werc: ooe unss on e a. , w wouee. unreanransorme sranave rseo coonormn uns on com sormeos no,emonsran successu sureesmsoernaon. oreee coesoonnesnromee sosnecawa wnroucon oe auos o wou an roe on aurcucmessnu a see °conncuaaeo wr.erenocuaeno m eours,e nm o eac cuure waseermne, aneasms werer ae erxoxramceaerom maunvuaaecnure vro’sumreos o eac cuure w a enasm exracon accornoe asms wereueocoo. nasm, m concenraons were measure usn aunsrumen.oae ono an xe:. woane, an o eac same was u araroseeor eecroorec anass. row. reee coo snoewssrewe s cueccesesnus anaene a eac cuuren m aer o asm emnaee sence o ,emonsranaema ure soowss srean’assm auexosron. row. ree coonesroucemaearsm m, oe cmuonusreraonr eaca coeon aer o r*ene exresson casseene muan as-conannos conerse anseroon su oore recaon o conrmneue erxem ec sa eow szsas e am oaro. eseaesms o r seacmoves oasms exraceromese cuures, xame:nuce conro o exresesreon. or n sran carr reesu aceeorasc acn c ene w corresonn ourer on neoro rec o ss reasnsas ceamn ccaesseeenee c aonnenoesennucnser oonraessmen nvoernnoer:oecsse e ao..: - - anro aurceoens,crouer anaonae.manoes ca sserasn os wnere consruce vaomooous recomnaon snae conanenomc conex.n aen ca aser o m reocceaneon rs,encae s: an rove ann nuceoe eacemen, ran s oenomc conexorenserons. ame errroavmen ocaon aernee neeureancee : meos, anace o eeecroocuormoesen sr sarnas was mae eecrocomeen accorno sanar onuraon orasm urenssso wrereran,soernm seeec we onmaauscm mea aaraes wou arnne.ransormaon wasm onuraoonse m resnumea norneoceo.: - -nv r ceon eoo nnr: e aense osns r a e a se. *ecenoen exareesss,onn ccaassene suncceess neuwuonscs,on o eacenomcanerae ene exresson.s suese aove,eacrouneve onc euxrnessoonser wnucer romoers was suceno suora omenuce a sm recaonneseoss.ransormaon w ossemrvaonsnuressoraemons resuen coones onnoss-, smarorevous unmuaer raeac o coma w ceranasms ane ree cooneassmen romroe. eurcanosno armnaoen oae om eaacnoses ser sarnanrsan wsasnaese asoows. onuraon werenocuaen orme w maasm an roe o m auos o mnma meum wou arnne cuure was one aermrunme,n a ane °anscmuasor.er aroxmae ,e nm o eac w a enasm exracon w acecreor exnraceoroem euvaen voumes o eac cuure concenraons were measure usn aunsrumen. manuacurer’srooco.asm :. woane, an o eac same wasoaea osnmo asn w aeraeroseueeoor ee n, mxe anass. croorec examne an a secon exermen,enuc oasmrouconneseoss was rsanosoorwmse. wne comoanesasrmomeransoormnauonaes oos an auos o roooss raon werenocuaeno m , an roe on arum, ro wm ramnose , orm eermne, aneasms werne a exra °cen acnua anoar.ze.er aroxmae ,e was nvoernnoer:oecsseo.: - - orowu e nre a. m s.ows sera cens reeancse a a samnear ce eacen sse o,n cucuarensn su mccnesmsau m reeac aer an reenon oe aon urem saowsasme. nucn a ceens aane eac cuurenm o or arnne,eeno as serrans o nrow. cuuresrew successu,uvena conans e a o snesze enoeono nuese aron renea cneasmorow.owever,sossea avanae an seeconressureor reenono onerre emaasm reresens arow conon.ucucree sowseasmesere ma osm cu.ureor eac sran an eacrow oss wensuscueasrm*roeucon was oserverom a srans an conons, ween mnma mea comareoe onee cronwsorucosssrouce aroxmaewceeasmewce as mucasm wennucen coma.reeoseweo unonss asorouce aroxmae mnma meau.re seowso assarosoesses- o, sa wmces c oonaasnmes eexurcaece conromo cnu.uresrownn reomnan sneasma r* muanene, sow a seces w a sncannarse,ro msoec suoawrs weoeasman ae exece sze an escrenx . ss conssen w enns mea wua orm uee an s. wowsn auarconse aeesn os. samasmes o szeass amre ass e exxraeccee arom cuuresrownn asescreor mnma mear n sow sneans . ow.ss conssen wennsescrenxame nvoernnoer:oeceo.: - - xsasm e ae.:euce ormonaeasmumerormao conannasms vrue o oma sr nn an can ueoor recaon o some,u no a, or- xame, as a sonaneouesr m counasouve exresson oerene.sescren seuence sac nnsrener*aroaenseasm aesmama,os srans can suor. n .asm a wasraasnmsorm coenannoneman oracone, was orna creae cou sowes re an acaareoseeo w sraens u anncu waasn ceom,eae we unmeruoaeeerrm-neenee.asmure asmreara ane anac-eseane, r ane arrow. onaon,aenssn aaeea ars aoasm .e correc szeasmsncae w an reresenn mume vee exece szene uneserearaon,e coones ourers . s. noewsemaeme,aesesn oaeasm an arrow, recovereromransorme exece sze an onem aonaan, was useoransorm woca. asr sownann aue u,n uncou aanve uneseasmrearaonsr re o omsransormaonnce arrows were w aasr souano aeee exece sze an onanarer moecuar we ans. mumersane oer anrmne:so eaeorn o an rera onrsor s moreeoenerae monomeranves rseo m maon o aasm monomeran oonomer-on w coones. ransormo an a asa exnc,s reesuromnenee sneownsnure an an useo. nvoerneoceo.: - - aaercno osecra:e e,ss ane e un a. rceae ac mcoornonmeroc saanna oreacsmnu weox was excserome srane wasra es.n auo o comeen ces o eac ae w mnmansorm mee aacc wornuoco ssanar meos, anransormans were seece on a rown overnn mnma mea we.ucxoseee.n coasomness w weerreeucreerom eacae an w sanar meos an run on an romese cuures -s ranso aaroseeo vsuazeeasms.ure - asmresromrmeno more.reueunre so -w more n -umserousarnanssoramneono . . serownn reresenaves warosmenur soesuoeermne arer moecuar we ansn seec an sown muean asn werer aeraeena wcasm exo.nueceraessee,n wavcereessrom onear , or resrcon enzmeco, accornoe manuacurer’snsrucosns ncenezm anes oaneerroman.s.acco enzme-reaere was run on an aaroseeo cecorerssence sownnure. reerenecseoon souure res,u eacn a se sn oenearrouc.e resus are ese or co ese, resecv ree coumnss uncu enomc an a nceasm e.-reaeresrom ncaea owever,nwo oeour sames o accouno,re aoes noon eamansresenne uncu sames.ncaeasm mumer naee exraans.e arrows e exer ans unaece exonuceaseanes an. o sran resumennsronaecra seeueanc neo co omnoese sonaneous muaonnerene aren sran, ,u asoensa noo acernoesrsa vearoaussmasm mus cmoemrs.areosss norneoceo.: - - av eve reanno nar: oeo auesss e “smeac aau. ”sea msummerasc aroenea.reru anm reercucea osrm esmnaee avanaesescrenseronsn euarocoss, wc ma creaensa. ma, w aso reesuu-n concaemer ene,oesenoeneraeasm mume en r- xame:nr uaneners.uorsecaon oma co asms, eoancu asm ow- eraarnon one oree one oree aees oerenervenransorme weeremnaromaoers anan ressanceaoa o nneasms, were co- on mnma meaaes smucose woannamuracno.nnso sr.a co ar* srans anae so coonne conona essenaene ar an w norown oans m annnmrao muceemer e suressor enes exresser a uness noe reca omemaasm.urer,emaasm w asms.eecanon oe asmenace oas amuncona warserneeroucromeow-co eenomcnseron cassee exressner* muavnouseneeuer nmoneoe suore eree aees oerenencuee co er aee. r ene w annseron : orr*en-ur ene :, enruncae :.ereer e, anrene we,u :,-osm :omoersc usencueer w eene amnose :. anuce a saonarase, an- oserve wene reesurs o*esn eexse erxmreensse arero smow eancn oeureree.rom.o coerransormans are oerwo aees sowsransormans.us,eeneroucs oerse,neu an neeer ore nvoernnoeoceo.: - - eansem wr:eses e, a.uenruncae, are unaeoacaeasm recaon oema xame:.ecomnanroenxressonn. conesence onocsor roen exresson was assesseeecon seescmons n an anoc-ree vecor ssem.eassor a e ar* amer so muanne enome, wcs comemene e anamcn caussereesssor n eacesasm.onro vecors ree on sanar er owooern seesecon. nuceromoer aser or ase was exresseromwo orn o sra or -nuce nasms w eere or a rencaon. anoc-reeeun eec xs ee renscsoronmo eere -ruaosnerucon reorer or ase were conenoe a romoer oro ann vecor- wnuc eeer aromo orer, or orenen oes re wercea coon an conann a exresson vecors conann aanamcn ressance cassee as conros, accornennoo saanear. norneoceo.: - - *veno a -r:e-ss e: au.cose,cero, ramnose, ,a , , ,a , *s*raacreae-,eaas:,eu acmos,neo, aacces,ron,n,o,u, ,o,e, , aeas, ameno, acas,n,,nacos,e,o,u,a, ,,o,,e,,a, as,arraacee, comeenacas omr conancnonroe coenaeeoeese wasransormeno cemca cemcaene non-nucn aar mean wsruocus ar.n cone ces vaea soc anae on on anamcn.aes werencuae overn a or an °o.csnceon coros were seeceransormaon was seece. onrom eac meum woeu se arecnene co oron an waoscrsowcnonnroos w.ere mro own c wemcaanamecnne a nnon-nncuucn overn a °.e overn cuures weren aenucon mea wou ocuaeno. m oeene auo cuures wererown a a °rnorne o.r aneocsconros wererown wanamcn.e eees werese an cen creus weereo saervaersaeeo cenruaon anrozen. an soueroenracons wereenaure an run onoaacr anam soueracons.oa conons. ee uner reuce ensomeroo anmaassses-s oaneroenaness w reeraev sceanone s,an anar esx oressnoonwneverso weenre co ensmae ommerca avaaeemae soware was useouan enens o oservceenraanons. araonn sannnenssneren w se oro . are sownnure. en sann.e-ae resus nvoernnoer:oceo.: - - -nuecss e anoc-reeen eee a. xs ernescsorom onno ve eer corenr was – eeranua er a renoru oec rr oorn orn o rase were conenoe an -nuce romoer, oreenes were co ecaon an conann anamcn ressance cassee as conros, accornoaneen.o conro vecors conann a comeenacas omr conancnonroe coennseroeese wasransormeno cemca cemcaene non-nucn aa ucs. co ces vaea soc anae on onormaanoa r mea wou arnne or anocsconros were seecerans nmcn.aes werencuae overn a °.ne coonrom eac °conroe w s s weas eerc se eeec ro coe wno.n wes weraenamrowcnnn.. m o meum anncuae overn a m o meumconros we e overn cuures werenocuaeno. werencuae a rerown wanamcno a sarn600o..uures cuures werencuae ° o uvnern an a600° o..e wsas w reeraecaerve asnenu cceenr wuao mn anroz.en.e nvoernnoer:oecseo.: - - an souees e roee a. nesra wceroenss weere ane cneanurreu aeno ru snea ornaeooaacr anam soueeer uanceorns r.euoa conons. ce en oomasse sane es were scanne, an exressoneves were esmae omsmoemrcear a anass oroenans reaveo sanars onownroen concenraon. an, varaonvan saaennenmeanse soware was useouan enens o oserveans. resus are sown sneren w se oroen sann.e-ae ramnosesee se an cuononur aruecs caornons an s ounsrae. em, eonserra oe moarnea romoer ssem werene aroraea ua or auonucon woue suae anuanucon acnrouunore ssrcaenn. anoc-reeen eexs erenscsoonn ve eercaeser reorer or ase were conenoe a romoer or ancor- wnuc eeer arom oorer, or orene one recaon an conann a vecors conann aanamcn ressance cassee as conros, s were coneno conro acasm conanneene accornoae orae. comeen oeese wasransormeno cemca cemcaene n oorn-nucn acoanr mroe coan wsruocs. co ces vaea soc anae on on anamcn.aes werencuae ovuer anrnne a or an °o.csnconros were seeceransormaon was seece e coonrom eac anamcn aen seneccuea ceoo. onveersn were arow °n.n. m o o overn me cuumucreon wraossn woecrueareownn wo nvoernnoer:oeceo.: - - reca oes oss ... e u m a. ure os were mnecuum w no seeconconros wanamcno a sarn w600m or m ramnoase a ° un an600o. was reace annuce e recaes we e as arorae. recae cuures werenuce, an e cerse w noen-nuce.e cuures werencuae overn a °.n rearvese cenruaon, wasewce w waer, an resusene was measu wraee.r.e rem oan rensus ceens weer ceees weeere arnansreorzreeno a weae, anuorescence e, anuoere csecenceee wsas w mereeasusree. on ao oecuoaarsor. a eevc seasmeec wrasranserreo a we assessaser- exresson.xcaon: nmms aax ae reaero sows aarra sown averae oese cesosn: nmuo: nm.ure ase ese exermensemonsrae exresson owo.enesaser an orromw oonuceromoersra or -nucenasms w eeren rn o recaon, wereeassor seeconse ar* amer so muan eaecasme.nome, wcs comemene easm uressor carre

Claims

nvoernnoer:oecsse e ao..: - - .as ca eerooourm soeaorso: nucc reeocome cnoamnos ce,eroaroc recomnanos ce exressn a unerone arames muaon,rsn an oen reanrame comrsn areneaas ownsreamne reanrameerames muaon resunn aremaure so coon conor aruncae recaon,neaorameserene oeraneo aromoer an . eroaroc recomnanorsroen. ram ce o cam, werenereneroroe aenensens meuaonse w erene, an werenerames muaon comrses an :ron onoe weo semeneweenosons an numere reaveo . erene. ramese muroaaornoc recomnanos ce o cam, werenereneroroenseronnoeoser ene, an werenerames muaon comrses an aenne : oer- e snemeneweenosons an numere reaveo . eroaroc rece. . omnanos ce o cam, wereneremaure so coons .rene weroear aoencn reecnosmernoannos ce o cam, werene seuence oeramesen.eraeenoroaeroenocm reec oomenarn com oaorsrs o ce ces r o cam:, w.erenerameserenes . eroaroc recom ecomnanos ce. os cesecenne recneanne.os ce o cam, wereneroaroc recomnannvorneoceo.: - -n.enor: eraeees norso ea a er. reocc reencoem onaenenos ce o cam, werenerameserenes . eroa ome oeroaroc recomnanos ce. roen comrsesroc recomn:an,os, o ce o cam, wereneruncae recaonnaor susuon, or a co r, or a varanereo comrsn a conservave . eroamnaonereo. coesor a recarooncn recaomornanos ce o cam, wereneramesereneurer com.rseseroaroc recomnraoneno.s ce o cam, werene recaonnaorroen . eroaroc: reco omr a vnaarnanose creeo comrsn a conservave susuon. comrses :, a varanereo o co cmarmsn, a w coenresenrve recaonnaorroen seecerom , , , , ave susuon, a muaon or-, or a comna , , , aeeon o amno acs- o.s ceeuroaroc recomonnanereoos. ce o cam, wereneroaroc recomnan comrsn arerr exeneresses aeerooousonuceoe comrsn an oen reanramenaorroen. ,erene oeraneo aromoer an conor a recaon o.nuceoeeroaar eoxrcess reescomernaamnesose cere one a cnamee,er wereneeerooous exresseserene ar ooousonuceoea . eroaroee sameeerooousonuceoe. onuceoea exrcess reescomernaamnesose cere o cam, wereneeerooous exresseserene areereneerooousonneuc aneoees.eerooousonuceoeanvoernnoer:oecsseo.: - - w. ero ea ar.oc recomnanos ce o cam, werenerene comrsese r.eerreonea.roc recomnanos ce o cam, werenerene comrsese com.en rsesee.roaroc: reco omr a vnaarnos ce o cam, werene recaonnaorroen . eroaroc recomnaanereo comrsn a conservave susuon. comrses :, a varnanose ce o cam, werene recaonnaorroen seecerom , , , r,eo com,rsn a conservave susuon, a muaon or-, or a , , aeeon o amno acs- . eroaro cocm recnaonereo. eenome oeroaroomnanos ce o cam, werenerenesneraeno n.eoeerroamaersoce recormc rec enoamnnoasn ceos o ce ca.m, wereneromoeras oera . eroaro nes annuceromoer. neoeramesce recomnanos ce o cam, wereneromoeras oera ara ,o,e-on orre,nesn one oamnosera , ac ssem,ranose ur.er comerroseasr aoc recomnanousc cee, o or camn,u wceere.neeerooousonuceoeer.mnaore cormoarrsoer ncm rencaoomr.nan:o.s ce o cam, wereneermnaor comrses anvorneoceo.: - - o.enor: s ce eees xrs ro e essa a er. so ac reeecroomonoaunsoosn cueceo o cea cmom,r wsnere an coenroonaaro ec recomnan comrsn ssenaene o.nuceoe a ereoaara eon xrc m ess ru eescaomon. ernaamneos ce o cam, wereneeerooous exr serene aneeerooousonuceoea .essesee croonaroonac re ecssenaene aree sameeerooousonuceoe. onuceoea exressesomernaamnesose cere one a cam, wereneeerooous exressese conona essenaene areerenenerooeoeuerooousonuceoea com. eroaroc recomnanos ce o cam, werenseo connucoenoaes e.ssenaene com.rses rsese an ero on araer ooece receonmesnsaneons caee os c oarm., werene conona essenaene com.rseseroarocne re.comnanos ce o cam, werene conona essenaene com.rsesee anr a aosareonec. recomnanos ce o cam, wereneean muaon a. eromarero mcu raecoonm.nanos ce o cam, werene amer muaon comrsesru. so ncae rea csomon cao cn nomac nre as o on a r:e. roen an ex orresnse o reecaroonaras caae o acvaon e ,e recaonnaorroen exresse eroaroco recco rmecomnannanos ce o cam e recaonnaorroen exresse eroaroc recomnanoso cse ce o o ca cmam, or,nvoernnoer:oecsse eo.: - - an coemona reounaoe a rre.o seuenc ae sse aenca exer marer an aeerooousonuceoe oera . esseneroaroc recomnanos ce o cam .. eeaassmm o o c caamm,, w weerreennee o orrnn o o r reeccaaoonn sesuenc-ee cromver.ses an o .s: ore-as omr o- c.a:m., werene orn o recaon comrses : e.eroooeusaosmnuc oeo caem comr wsnere an coneroonaaro esscen recaomennea cnomorss ce exresses a muaon, ane seecae marer comrses a srucuraene seue n aean e suressor comrsn an ancoon caae o comence o a suressor , ess menne conona e.ena eroooeene. usaosmnuc oeo caem com,r wsnere an coneoronaaro esc recomnanos ce exresses a muaon, ane seecae marer comrses aus senaene comrsn aean conon euenceeneo comemene cas. a see easssmen oa ceanem., werene seecae marer comrses an anoc ressance .e.e:as om o cam, wereneene seuence oe suressor comrses . ear :. o aromoer.sm o cam, werene suressor ene seuences oeranenvorneoceo.: - - .enor: eneeess seas e um a. ence o co cmarms, wereneromoeras oeraneoe suressor . easm o cames, werene: su orre :. o aerm ssor ene seuences oerane . n eneeaor sea.sumen o cam, wereneermnaoras oeraneoe suressor . easmce o co cmamrses, wer : or :.ne eneeerooousonuceoe coesor aroen o a.res. ransosaese,asmroe ons coarm vra, v weceoreneroen oneres comrses aeraeucroen, a vecors, anroensrouce recomr snessncun vraacanroens anranser nurona suemenaon,nusrananoroo, cosmecs,oues, arcuuraee, an a. resrcesaesmas om c’as cmom,a were cnem oecaas,sm an cuornsuemr ceormoorsse.s anncoma acor . aos cea exresseserameserene. cuvan meeoroaorro muc recocmaonnan ooeasm o cam,e meo comrsn: roaroc recomnanos ce seesc cne oe caamsmromnroeu ccen ceasmnoe en carre oun a meumree o a nuren vaoreroaroc recoumure,e seecon row an cuvane seece ce nanos ce’s .. ee m meeoo o o c caamm,, w weerreennes e e. exxreressssoonn o oeerarmameseseerrenees consuve. . e meo o cam, werene exr nesnuce.nuce. esson oerameserenes auo-nvorneoceo.: - - mu.enor: n meess e eo a. aosmrro ou ccanm a recomrnanroen oneres,e meo comrsn: comrs ansormn aos ce easm,eos ce an an:eero aoeoerooousonuceoe comrsn arene oeraneo aromoer, eos cen a cuuusre moenuucmeo ae comrsn a conona essenaene cuvan urne recomnanroenow onne erxresson oe recomnanroen oneres an os c esrome cuure meum orromeransorme ca.e. m ae meo o cam, wereneos ceseroaroc recomnanos ce o . ne meeor oene caserameserene. ce m, wereneos ces a seconroaroc recomnanos .. . ee m meeoo o o c caamm,, w weerreennee e exxrreessssoonn o oeerreennees consuve. . e meo o cam, werene exre snuce. . e meo o c sson oerenes auo-nuce. romoer ane exressaonm o,e wreorene exresson oerenesnuce w one . e meo o cam, werenenes exnruecssoen w oe areerenne anromoeer. we sa roensnuce w.e mm aseeoromoorerr.earn a moe-ce,e meo comrsnransecne-ce ce. memo oor caremarn. vra vecors,e meo comrsnran m wrseseenesa esnmcon o e ceammens r,e w secn a euarocos co ureerenorreouecenro voroaus vecoorsnunceoe euea oroceosas cme.nvoernneoceo.: - -.or: meesso e ao.r use oeasm o cam as aemaeorn vroranscron.

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